Instrument: Illumina HiSeq 2500
Strategy: ncRNA-Seq
Source: TRANSCRIPTOMIC
Selection: size fractionation
Layout: SINGLE
Construction protocol: Cells were collected by trypsinization and washed in PBS. Cytosol was isolated by plasma membrane permeabilization and differential centrifugation. Cytosolic RNA was isolated using Trizol LS. Indexed RNA-seq libraries were made using the Illumina TruSeq Small RNA kit. Libraries were amplified with 14 cycles of PCR, pooled, and separated by PAGE. Products from small RNA with input sizes of 35 to 350 nucleotides were excised, collected, and sequenced using an Illumina HiSeq 2500.